Immunolabeling of Cleared Human Pancreata Provides Insights into Three-Dimensional Pancreatic Anatomy and Pathology

Michaël Noë, Neda Rezaee, Kaushal Asrani, Michael Skaro, Vincent P. Groot, Pei Hsun Wu, Matthew T. Olson, Seung Mo Hong, Sung Joo Kim, Matthew J Weiss, Christopher Wolfgang, Martin A Makary, Jin He, John L Cameron, Denis Wirtz, Nicholas Roberts, G. Johan A. Offerhaus, Lodewijk A.A. Brosens, Laura Delong Wood, Ralph H Hruban

Research output: Contribution to journalArticle

Abstract

Visualizing pathologies in three dimensions can provide unique insights into the biology of human diseases. A rapid and easy-to-implement dibenzyl ether–based technique was used to clear thick sections of surgically resected human pancreatic parenchyma. Protocols were applicable to both fresh and formalin-fixed, paraffin-embedded tissue. The penetration of antibodies into dense pancreatic parenchyma was optimized using both gradually increasing antibody concentrations and centrifugal flow. Immunolabeling with antibodies against cytokeratin 19 was visualized using both light sheet and confocal laser scanning microscopy. The technique was applied successfully to 26 sections of pancreas, providing three-dimensional (3D) images of normal pancreatic tissue, pancreatic intraepithelial neoplasia, intraductal papillary mucinous neoplasms, and infiltrating pancreatic ductal adenocarcinomas. 3D visualization highlighted processes that are hard to conceptualize in two dimensions, such as invasive carcinoma growing into what appeared to be pre-existing pancreatic ducts and within venules, and the tracking of long cords of neoplastic cells parallel to blood vessels. Expanding this technique to formalin-fixed, paraffin-embedded tissue opens pathology archives to 3D visualization of unique biosamples and rare diseases. The application of immunolabeling and clearing to human pancreatic parenchyma provides detailed visualization of normal pancreatic anatomy, and can be used to characterize the 3D architecture of diseases including pancreatic intraepithelial neoplasia, intraductal papillary mucinous neoplasm, and pancreatic ductal adenocarcinomas.

Original languageEnglish (US)
Pages (from-to)1530-1535
Number of pages6
JournalAmerican Journal of Pathology
Volume188
Issue number7
DOIs
StatePublished - Jul 1 2018

Fingerprint

Pancreas
Anatomy
Pathology
Pancreatic Neoplasms
Paraffin
Formaldehyde
Antibodies
Adenocarcinoma
Keratin-19
Pancreatic Diseases
Venules
Three-Dimensional Imaging
Pancreatic Ducts
Rare Diseases
Confocal Microscopy
Blood Vessels
Neoplasms
Carcinoma
Light

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Immunolabeling of Cleared Human Pancreata Provides Insights into Three-Dimensional Pancreatic Anatomy and Pathology. / Noë, Michaël; Rezaee, Neda; Asrani, Kaushal; Skaro, Michael; Groot, Vincent P.; Wu, Pei Hsun; Olson, Matthew T.; Hong, Seung Mo; Kim, Sung Joo; Weiss, Matthew J; Wolfgang, Christopher; Makary, Martin A; He, Jin; Cameron, John L; Wirtz, Denis; Roberts, Nicholas; Offerhaus, G. Johan A.; Brosens, Lodewijk A.A.; Wood, Laura Delong; Hruban, Ralph H.

In: American Journal of Pathology, Vol. 188, No. 7, 01.07.2018, p. 1530-1535.

Research output: Contribution to journalArticle

Noë, Michaël ; Rezaee, Neda ; Asrani, Kaushal ; Skaro, Michael ; Groot, Vincent P. ; Wu, Pei Hsun ; Olson, Matthew T. ; Hong, Seung Mo ; Kim, Sung Joo ; Weiss, Matthew J ; Wolfgang, Christopher ; Makary, Martin A ; He, Jin ; Cameron, John L ; Wirtz, Denis ; Roberts, Nicholas ; Offerhaus, G. Johan A. ; Brosens, Lodewijk A.A. ; Wood, Laura Delong ; Hruban, Ralph H. / Immunolabeling of Cleared Human Pancreata Provides Insights into Three-Dimensional Pancreatic Anatomy and Pathology. In: American Journal of Pathology. 2018 ; Vol. 188, No. 7. pp. 1530-1535.
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AU - Groot, Vincent P.

AU - Wu, Pei Hsun

AU - Olson, Matthew T.

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AU - Makary, Martin A

AU - He, Jin

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